|Detection of Human IL‑2 by Western Blot. Western blot shows lysates of monensin treated human peripheral blood mononuclear cells (PBMCs) with no additional treatment (-) or additionally treated (+) with 0.5ug/mL calcium ionomycin (Iono) and 50ng/mL PMA overnight. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL‑2 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|IL‑2 in Human PBMCs. IL‑2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with PMA, ionomyocin, and monensin using Goat Anti-Human IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.|
|Cell Proliferation Induced by IL‑2 and Neutralization by Human IL‑2 Antibody. Recombinant Human IL‑2 (Catalog # 202-IL) stimulates proliferation in the CTLL‑2 mouse cytotoxic T cell line in a dose-dependent manner (orange line) as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Human IL‑2 (2 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-202-NA). The ND50 is typically ≤0.15 µg/mL.|
Interleukin-2 (IL-2) is a O-glycosylated, four alpha -helix bundle cytokine that has potent stimulatory activity for antigen-activated T cells. It is expressed by CD4+ and CD8+ T cells, gamma δ T cells, B cells, dendritic cells, and eosinophils (1 - 3). Mature human IL-2 shares 56% and 66% aa sequence identity with mouse and rat IL-2, respectively. Human and mouse IL-2 exhibit cross-species activity (4). The receptor for IL-2 consists of three subunits that are present on the cell surface in varying preformed complexes (5 - 7). The 55 kDa IL-2 R alpha is specific for IL-2 and binds with low affinity. The 75 kDa IL-2 R beta, which is also a component of the IL-15 receptor, binds IL-2 with intermediate affinity. The 64 kDa common gamma chain gamma c/IL-2 R gamma, which is shared with the receptors for IL-4, -7, -9, -15, and -21, does not independently interact with IL-2. Upon ligand binding, signal transduction is performed by both IL-2 R beta and gamma c. IL-2 is best known for its autocrine and paracrine activity on T cells. It drives resting T cells to proliferate and induces IL-2 and IL-2 R alpha synthesis (1, 2). It contributes to T cell homeostasis by promoting the Fas-induced death of naïve CD4+ T cells but not activated CD4+ memory lymphocytes (8). IL-2 plays a central role in the expansion and maintenance of regulatory T cells, although it inhibits the development of Th17 polarized cells (9 - 11). Thus, IL-2 may be a key cytokine in the natural suppression of autoimmunity (12, 13).
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